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TitleStructure and function of H/K pump mutants reveal Na/K pump mechanisms.
Journal, issue, pagesNat Commun, Vol. 13, Issue 1, Page 5270, Year 2022
Publish dateSep 9, 2022
AuthorsVictoria C Young / Hanayo Nakanishi / Dylan J Meyer / Tomohiro Nishizawa / Atsunori Oshima / Pablo Artigas / Kazuhiro Abe /
PubMed AbstractIon-transport mechanisms evolve by changing ion-selectivity, such as switching from Na to H selectivity in secondary-active transporters or P-type-ATPases. Here we study primary-active transport via ...Ion-transport mechanisms evolve by changing ion-selectivity, such as switching from Na to H selectivity in secondary-active transporters or P-type-ATPases. Here we study primary-active transport via P-type ATPases using functional and structural analyses to demonstrate that four simultaneous residue substitutions transform the non-gastric H/K pump, a strict H-dependent electroneutral P-type ATPase, into a bona fide Na-dependent electrogenic Na/K pump. Conversion of a H-dependent primary-active transporter into a Na-dependent one provides a prototype for similar studies of ion-transport proteins. Moreover, we solve the structures of the wild-type non-gastric H/K pump, a suitable drug target to treat cystic fibrosis, and of its Na/K pump-mimicking mutant in two major conformations, providing insight on how Na binding drives a concerted mechanism leading to Na/K pump phosphorylation.
External linksNat Commun / PubMed:36085139 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.8 - 3.4 Å
Structure data

32954

7x21

EMDB-32954, PDB-7x21:
Cryo-EM structure of non gastric H,K-ATPase alpha2 K794A in (K+)E2-AlF state
Method: EM (single particle) / Resolution: 2.8 Å

32955

7x22

EMDB-32955, PDB-7x22:
Cryo-EM structure of non gastric H,K-ATPase alpha2 K794S in (2K+)E2-AlF state
Method: EM (single particle) / Resolution: 3.0 Å

32956

7x23

EMDB-32956, PDB-7x23:
Cryo-EM structure of non gastric H,K-ATPase alpha2 SPWC mutant in 3Na+E1-AMPPCPF state
Method: EM (single particle) / Resolution: 3.2 Å

32957

7x24

EMDB-32957, PDB-7x24:
Cryo-EM structure of non gastric H,K-ATPase alpha2 SPWC mutant in (2K+)E2-AlF state
Method: EM (single particle) / Resolution: 3.4 Å

PDB-7x20:
Crystal structure of non gastric H,K-ATPase alpha2 in (K+)E2-AlF state
Method: X-RAY DIFFRACTION / Resolution: 3.3 Å

Chemicals

ChemComp-ALF:
TETRAFLUOROALUMINATE ION

ChemComp-K:
Unknown entry

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

ChemComp-HOH:
WATER

ChemComp-MG:
Unknown entry

ChemComp-CLR:
CHOLESTEROL

ChemComp-PCW:
1,2-DIOLEOYL-SN-GLYCERO-3-PHOSPHOCHOLINE / DOPC, phospholipid*YM

ChemComp-ACP:
PHOSPHOMETHYLPHOSPHONIC ACID ADENYLATE ESTER / AMP-PCP, energy-carrying molecule analogue*YM

ChemComp-NA:
Unknown entry

Source
  • rattus norvegicus (Norway rat)
KeywordsMEMBRANE PROTEIN / P-type ATPase / transporter / proton pump / colon / kidney / airway

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